One of the biggest innovations in human prehistory was the advent of food production, consisting of the ability to grow crops and domesticate animals for consumption. This wide-scale transition from hunting and gathering to food production led to more permanent settlements, and set in motion major societal changes. In western Eurasia, this revolution spread from the Near East into Europe, Africa and diverse regions of Asia. Agriculture was brought into Europe by the descendants of early Anatolian farmers starting approximately 8,000 years ago. But little was known of the people who developed agriculture in the Fertile Crescent: where they all closely related to the early Anatolian farmers, or were there multiple ethnic groups who developed agriculture in parallel? In the first data chapter, I use the first genome from a Neolithic woman from Ganj Dareh, in the Zagros Mountains (Iran), a site with evidence of early goat domestication 10,000 years ago. I showed that Western Iran wan inhabited by populations mostly similar to Hunter- gatherer populations from the Caucasus, but remarkably, very distinct from the Anatolian farmers who spread the Neolithic package into Europe. While a degree of cultural diffusion between Anatolia, Mesopotamia and the Zagros highlands likely happened, genetic dissimilarity supports a model in which Neolithic societies of that area were distinct. The second chapter deals with how Africa was affected by population movements, originating in the Near East, during the Neolithic times. Characterising genetic diversity in Africa is a crucial step for analyses reconstructing human evolution. Using Mota, an ancient genome from a male from the Ethiopian highlands, I showed a backflow into Africa by populations closely related to the Anatolian Neolithic farmers. The third chapter deals with some common problems and themes in the analysis of ancient DNA, such as merging capture datasets with diverse number of ascertained SNPs, combining capture and shotgun data in the same analysis, and the effect of UDG treatment in ancient samples. I describe the most common problems and their effect in summary statistics, and propose a guide on how to work with ancient DNA to avoid data compatibility problems.

The Quaternary (the approximately the last 2.6 million years) of North America is a tremendously exciting time period to study with respect to ecology. It saw periods of immense climatic turbidity - the expansion and retreat of continental ice-sheets and large swings in temperature, resulting in the wide scale restructuring of terrestrial ecosystem. It also saw widespread migrations of many species and out of Eurasia, mostly notably of modern humans. Ancient DNA offers powerful tools to examine the relationships and responses of megafunal species to these events, but has largely focused on cold-adapted species, and within radiocarbon-time (i.e. the last 50 thousand years). In this thesis I work to expand our understanding of the genetic landscape of Pleistocene megafuna in three ways. First, I describe the analysis of coprolites from Bechan Cave, Utah and characterize the mammoth inhabitants in the broader context of North American mammoths. Second, I characterize the diversity of American mastodons across the continent and through time, showing that their range likely repeatedly expanded and contracted in response to Pleistocene glaciations. Lastly, I begin to fill in some of the gaps in the American mastodon dataset from chapter 3, and begin to address some of the taxonomic and biogeographic questions about American and Pacific mastodons in Idaho. Understanding how North American megafauna responded to these climatic and anthropogenic stresses may help to explain why so many species went extinct at the end of the last glaciation, and how species may respond to present day warming. However, it is important to include taxa from warmer locales and environments to ensure our models and hypotheses are comprehensive. / Thesis / Doctor of Science (PhD) / Pleistocene North America was a time period of immense climatic turbidity, with temperature swings greater than 15°C in response to the expansion and contraction of continental ice-sheets. Despite these massive swings in temperature, many species managed to thrive on the continent and adapt to glacial-associated ecosystem restructuring. Ancient DNA from Pleistocene megafauna can serve as a very useful tool to answer many questions about the distribution of megafaunal species, and how they may have responded to these climatic events. However, most studies have largely focused on species adapted to cold environments and from the last fifty thousand years. In this thesis I extend our knowledge of the genetic landscape of Pleistocene proboscideans, characterizing the mammoth inhabitants of Bechan Cave, Utah, and producing the first look at American mastodon diversity through space and time. This work increases our representation of warm-adapted specimens and characterizes the effects of glacial cycles on megafauna populations.

Tuberculosis has been an important disease throughout human history, shaping countless past populations. The archaeological study of the causative agents of tuberculosis, members of the Mycobacterium tuberculosis Complex (MTBC), is hindered by the non-diagnostic nature of tuberculosis-associated skeletal changes. As such, ancient DNA (aDNA) or palaeogenetic analyses have become an important tool for identifying tuberculosis in past populations. However, due to the age and variable preservation of aDNA, there are often issues with sporadic results and false negatives. The overall aim of the work presented here was to use different methods, including traditional target-specific PCR, to identify and detect tuberculosis aDNA in archaeological remains. The main objectives within this overarching aim were to first test a method called whole genome amplification (WGA), used to non-specifically amplify all the DNA within a sample, and its potential to improve the yield of aDNA from skeletal remains (Chapters 3 and 4). To determine the extent of its impact, WGA was used in a comparative context, where each archaeological sample analysed was separately subjected to two methods of MTBC detection - the traditional targeted PCR method and the same method assisted by the initial application of WGA. The results show that applying WGA before the traditional targeted PCR methodology to detect the presence of MTBC pathogens in skeletal remains is only useful and viable in some cases, likely depending on the age and preservation of the sample. The second objective was to use next generation sequencing to obtain more information on the aDNA composition of certain archaeological samples and answer questions beyond the scope of traditional target-specific PCR techniques (Chapter 5). Although most of the sequencing runs were variably unsuccessful, the composition of two samples, both known to probably contain tuberculosis aDNA, could be analysed. The samples both contained similar amounts of mycobacterial aDNA and varying amounts of both human and even potentially human intestinal flora DNA. Finally, the third objective was to determine if MTBC aDNA could be detected in a rib sample from Private William Braine of the lost Franklin Expedition using standard target-specific PCR (Chapter 6). In this case study, no evidence of tuberculosis ancient DNA was found. The work done through-out highlights the difficulties of ancient DNA research and, in Chapter 4, shows the importance of using more than a single sample to evaluate methods for application in palaeogenetic contexts.

La paléoparasitologie est l’étude des restes de parasites préservés dans des échantillons archéologiques et permet de mieux comprendre l’état de santé des populations anciennes et d’obtenir des informations d’ordre anthropologique ou ethnologique, sur les régimes alimentaires ou les conditions d’hygiène au quotidien. Les restes de parasites peuvent être retrouvés sous forme de macro-restes (vers ou larves), d’antigènes, d’ADN ou d’œufs. Cesderniers peuvent être particulièrement bien préservés au cours du temps car ils sont composés en partie de chitine, les rendant résistants aux processus de dégradation. L’observation microscopique de leurs caractéristiques morphologiques et micrométriques permet d’identifier les taxons au niveau du genre ou de la famille. Dans le cadre de cette thèse, plusieurs helminthes gastro-intestinaux, dont les œufs sont fréquemment retrouvés dans des échantillons archéologiques, ont été ciblés par une approche génétique. Il s’agit des vers plats Tæniasaginata, T. solium, T. asiatica, Echinococcus granulosus, E. multilocularis, Diphyllobothriumlatum, D. dendriticum et D. nihonkaiense, des nématodes Trichuris trichiura, Enterobiusvermicularis et Ascaris sp. et des douves Fasciola hepatica, F. gigantica, Dicrocoeliumdendriticum et D. chinensis.La méthode « aMPlex Torrent » permet de détecter, dans un grand nombre d’échantillons archéologiques, une faible quantité d’ADN de parasites. Cette approche combine la spécificité et la sensibilité de la PCR au haut-débit du séquençage de nouvelle génération. Plusieurs vestiges, provenant de périodes et de régions géographiques diverses, ont été analysés. Des résultats génétiques ont été obtenus pour des échantillons aussi anciens que 7200 BP. Nous avons par ailleurs obtenus les premières séquences anciennes de Taenia sp., Diphyllobothriumsp., Echinococcus sp., et les premières séquences européennes d’Enterobius vermicularis. Auvu de ces résultats, notre approche apparait comme étant complémentaire à la microscopie. / Palaeoparasitogy, the study of parasite remains from archaeological samples, is adiscipline that can highlight questions about the health status of the ancient populations. It can give important anthropological or ethnological information such as the diet and the hygiene conditions of past societies. The remains can be preserved as macroremains (worms or larvae),antigens, DNA or eggs. Because they are partially made of chitin, eggs of gastrointestinalhelminths resist well over time to the taphonomic degradation process. It is possible to distinguish between different families or genera of parasites by looking at the morphological features of eggs. However, since several taxa share common features, the determination is rarelypossible at the species level. For this thesis, several parasite species for which eggs arecommonly observed in archeological samples have been studied by a genetic approach. Westudied the tapeworms Tænia saginata, T. solium, T. asiatica, Echinococcus granulosus, E.multilocularis, Diphyllobothrium latum, D. dendriticum, and D. nihonkaiense; the nematodesTrichuris trichiura, Enterobius vermicularis, and Ascaris sp.; and the flukes Fasciola hepatica,F. gigantica, Dicrocoelium dendriticum, and D. chinensis.The “aMPlex Torrent” approach has been set up to detect minute amounts of DNA from parasites in multiple archaeological samples. This approach combines the specificity andsensitivity of PCR to the throughput of Next-Generation sequencing. Several samples have been analyzed by this approach. We obtained genetic results for samples as old as 7200 BP and from various geographical and archeological contexts. We obtained the first ancient DNA sequences for Taenia sp., Diphyllobothrium sp., Echinococcus sp. and the first European sequences forEnterobius vermicularis. Genetic analyses and microscopic observations appear to be complementary. Indeed, at least one taxon per sample was detected by one of the two approaches.

Humans evolved in a world of giant creatures. Current evidence suggests that most ice age megafauna went extinct around the transition to our current Holocene epoch. The ecological reverberations associated with the loss of over 65% of Earth’s largest terrestrial animals transformed ecosystems and human lifeways forever thereafter. However, there is still substantial debate as to the cause of this mass extinction. Evidence variously supports climate change and anthropogenic factors as primary drivers in the restructuring of the terrestrial biosphere. Much of the ongoing debate is driven by the insufficient resolution accessible via macro-remains. To help fill in the gaps in our understandings of the Pleistocene-Holocene transition, I utilized the growing power of sedimentary ancient DNA (sedaDNA) to reconstruct shifting signals of plants and animals in central Yukon. To date, sedaDNA has typically been analyzed by amplifying small, taxonomically informative regions. However, this approach is not ideally suited to the degraded characteristics of sedaDNA and ignores most of the potential data. Means of isolating sedaDNA have also suffered from the use of overly aggressive purification techniques resulting in substantial loss. To address these limitations, I first experimentally developed a novel means of releasing and isolating sedaDNA. Secondly, I developed a novel environmental bait-set designed to simultaneously capture DNA informative of macro-scale ecosystems. When combined, we identify a substantial improvement in the quantity and breadth of biomolecules recovered. These optimizations facilitated the unexpected discovery of horse and mammoth surviving thousands of years after their supposed extirpation. I followed up these results by extracting DNA from multiple permafrost cores where we confirm the late survival signal and identify a far more complex and high-resolution dataset beyond those identifiable by complementary methods. I was also able to reconstruct mitochondrial genomes from multiple megafauna simultaneously solely from sediment, demonstrating the information potential of sedaDNA. / Dissertation / Doctor of Philosophy (PhD) / A new addition to the rapidly growing field of palaeogenetics is environmental DNA (eDNA) with its immense wealth of biomolecules preserved over millennia outside of biological tissues. Organisms are constantly shedding cells, and while most of this DNA is metabolized or otherwise degraded, some small fraction is preserved through sedimentary mineral-binding. I experimentally developed new ancient eDNA methods for recovery, isolation, and analysis to maximize our access to these biomolecules and demonstrate that this novel approach outperforms alternative protocols. Thereafter, I used these methods to extract DNA from ice age permafrost samples dating between 30,000–6,000 years before present. These data demonstrate the power of ancient eDNA for reconstructing ecosystem change through time, as well as identifying evidence for the Holocene survival of caballine horse and woolly mammoth in continental North America. This late persistence of Pleistocene fauna has implications for understanding the human ecological and climatological factors involved in the Late Pleistocene mass extinction event. This effort is paralleled with megafaunal mitogenomic assembly and phylogenetics solely from sediment. This thesis demonstrates that environmental DNA can significantly augment macro-scale buried records in palaeoecology.